Biology subjects

Porcar, M.

Publications and source records attributed to Porcar, M..

12 recordsLinked to original sources

bioRxiv2023

Unveiling the ecology, taxonomy and metabolic capabilities of MBA03, a potential key player in anaerobic digestion

Biogas, a mix of CO2, CH4 and small proportions of other gases, is a biofuel obtained by anaerobic digestion (AD). Biogas production is often considered a black box process, as the role and dynamics of some of the microorganisms involved remain undisclosed. Previous metataxonomic studies in the frame of the MICRO4BIOGAS project (www.micro4biogas.eu) revealed that MBA03, an uncharacterised and uncultured bacterial taxon, was very prevalent and abundant in industrial full-scale AD plants. Surprisingly, no culturable specimen or genome of this taxon has ever been reported, so its role in AD has remained unclear. In the present work, thirty samples derived from anaerobic digesters were sequenced, allowing the reconstruction of 108 metagenome-assembled genomes (MAGs) potentially belonging to MBA03. According to phylogenetic analyses and genomic similarity indices, MBA03 constitutes a new bacterial order, proposed as Darwinibacteriales ord. nov., which includes Darwinibacter acetoxidans gen. nov., sp. nov. of the family Darwinibacteriaceae fam. nov., along with Wallacebacter cryptica gen. nov., sp. nov. of the Wallacebacteriaceae fam. nov. Ecotaxonomic studies determined that AD processes are the main ecological niche of Darwinibacteriales. Moreover, metabolic predictions identified Darwinibacteraceae members as putative syntrophic acetate oxidising bacteria (SAOB), as they encode for the reversed Wood-Ljungdahl (W-L) pathway coupled to the glycine cleavage system. This suggests that Darwinibacteraceae members work in collaboration with hydrogenotrophic archaea to produce methane in industrial biogas plants. Overall, our findings present Darwinibacteriales as a potential key player in anaerobic digestion and pave the way towards the complete characterisation of this newly described bacterial taxa.

ecology↗

bioRxiv2023

Multivariate comparison of taxonomic, chemical and technical data from 80 full-scale an-aerobic digester-related systems

This study represents one of the most comprehensive characterisations of the anaerobic digestion (AD) microbiome with 80 samples from 45 different large-scale reactors in three coun-tries. Technical, chemical and taxonomic data was thoroughly collected, analyzed and correlated to identify the main drivers of AD processes. Our results showed that MBA03, Proteiniphilum, a member of Dethiobacteraceae, and Caldicoprobacter were present in all the samples, while Meth-anosarcina was the most abundant and prevalent archaea. Two distinct bacterial clusters were iden-tified by correlating microbial abundances. One was correlated with hydrogenotrophic and the other with acetoclastic methanogenesis. Organic acids, ammonia, nitrogen, COD and the trace el-ements Fe, Mo, and the macro nutrient P had the greatest impact on AD microbiomes. Temperature, reactor type and substrate also influenced the formation of a specialized microbial community. Overall, this work sheds light on the microbial key players involved in AD and evaluates how they are affected by technical and chemical parameters. HighlightsO_LIGeneration of a holistic dataset of chemical, taxonomic and technical parameters of 80 large-scale anaerobic digestion systems. C_LIO_LIIdentification of a core microbiome comprising MBA03, Proteiniphilum, an uncultured or-ganism from the Dethiobacteraceae family, and the Caldicoprobacter family. C_LIO_LIIdentification of the main influencing parameters that determine the occurrence and non-occurrence of specific genera. C_LIO_LICorrelation of bacterial taxa with hydrogenotrophic and/or acetoclastic archaea. C_LI

microbiology↗

bioRxiv2023

Maltodextrin administration ameliorates brain pathology in a mouse model of mitochondrial disease

Mitochondrial dysfunction lead to a wide group of progressive and fatal pathologies known as mitochondrial diseases (MD). One of the most common pediatric representation of MD is Leigh Syndrome, affecting 1/40.000 births. LS is characterized by neurodegeneration in specific brain areas, such as brainstem and basal ganglia, and by respiratory and motor alterations. However, the results obtained from clinical trials based on antioxidant therapies are controversial. Thus, the development novel antioxidant strategy is required to improve the efficacy of current palliative treatments. In this regard, Ndufs4KO mouse model is a suitable model to test new drugs in the field of MD and LS. Therefore, we set to assess the therapeutic potential of oral administration of Micrococcus luteus, a high-antioxidant content microorganism. Incidentally, we identified that while M. luteus administration did not possess any beneficial actions, the cryopreservant maltodextrin (MDX), included in the preparation, ameliorated the phenotype of Ndufs4KO mice. Our results show that MDX treatment at a concentration of 30 g/L increased lifespan and reduced microglial reaction compared to vehicle-treated Ndufs4KO mice. However, no improvement in locomotion nor respiratory function was observed in MDX-treated mice compared to vehicle-treated Ndufs4KO mice. Metataxonomic characterization of intestinal microbiome identified differential profiles in Ndufs4KO mice at the genus level. Furthermore, MDX treatment increased the variability of the abundance of Akkermansia sp. Thus, this work paves the way for further studies to confirm the therapeutic potential of MDX in mitochondrial disease.

neuroscience↗

bioRxiv2023

A 3D printed plastic frame deeply impacts yeast cell growth

Solid State Fermentation (SSF) processes have been explored for yeast growth and protein and metabolites production. However, most of these processes lack standardization. In this work, we present a polylactic acid (PLA) 3D printed matrix that dramatically enhances yeast growth when embedded in liquid media compared to equivalent static cultures, and changes yeast expression patterns at the proteome level. Moreover, differences in sugar assimilation and ethanol production, as the main product of alcoholic fermentation, are observed. Our results suggest that these matrixes may be useful for a vast range of biotechnological applications based on yeast fermentation.

microbiology↗

bioRxiv2019

Out of the abyss: Genome and metagenome mining reveals unexpected environmental distribution of abyssomicins

Natural products have traditionally been discovered through the screening of culturable microbial isolates from all sort of environments. The sequencing revolution allowed the identification of dozens of biosynthetic gene clusters (BGCs) within single bacterial genomes, either from cultured or uncultured strains. However, we are still far from fully exploiting the microbial reservoir, as most of the species are non-model organisms with complex regulatory systems and yet recalcitrant to be engineered. Today, genomic and metagenomic data produced by laboratories worldwide covering the most different natural and artificial environments on Earth, are an invaluable source of raw information from which natural product biosynthesis can be accessed. In the present work, we describe the environmental distribution and evolution of the abyssomicin BGC through the analysis of publicly available genomic and metagenomic data. Our results demonstrate that the selection of a pathway-specific enzyme to direct the genome mining is an excellent strategy that led to the identification of 74 new Diels-Alderase homologs and unveiled a surprising prevalence of the abyssomicin BGC within terrestrial habitats, mainly soil and plant-associated, where we have identified five complete and 12 partial new abyssomicin BGCs and 23 new potential abyssomicin BGCs. Our results strongly support the potential of genome and metagenome mining as a key preliminary tool to inform bioprospecting strategies aiming at the identification of new bioactive compounds such as -but not restricted to-abyssomicins.

microbiology↗

bioRxiv2019

Assembly methods for nanopore-based metagenomic sequencing: a comparative study

BackgroundMetagenomic sequencing has lead to the recovery of previously unexplored microbial genomes. In this sense, short-reads sequencing platforms often result in highly fragmented metagenomes, thus complicating downstream analyses. Third generation sequencing technologies, such as MinION, could lead to more contiguous assemblies due to their ability to generate long reads. Nevertheless, there is a lack of studies evaluating the suitability of the available assembly tools for this new type of data.\n\nFindingsWe benchmarked the ability of different short-reads and long-reads tools to assembly two different commercially available mock communities, and observed remarkable differences in the resulting assemblies depending on the software of choice. Short-reads metagenomic assemblers proved unsuitable for MinION data. Among the long-reads assemblers tested, Flye and Canu were the only ones performing well in all the datasets. These tools were able to retrieve complete individual genomes directly from the metagenome, and assembled a bacterial genome in only two contigs in the best scenario. Despite the intrinsic high error of long-reads technologies, Canu and Flye lead to high accurate assemblies (~99.4-99.8 % of accuracy). However, errors still had an impact on the prediction of biosynthetic gene clusters.\n\nConclusionsMinION metagenomic sequencing data proved sufficient for assembling low-complex microbial communities, leading to the recovery of highly complete and contiguous individual genomes. This work is the first systematic evaluation of the performance of different assembly tools on MinION data, and may help other researchers willing to use this technology to choose the most appropriate software depending on their goals. Future work is still needed in order to assess the performance of Oxford Nanopore MinION data on more complex microbiomes.

bioinformatics↗

bioRxiv2019

Shedding light on biogas: a transparent reactor triggers the development of a biofilm dominated by Rhodopseudomonas faecalis that holds potential for improved biogas production

Conventional anaerobic digesters intended for the production of biogas usually operate in complete darkness. Therefore, little is known about the effect of light on microbial communities operating in anaerobic digesters. In the present work, we have studied through 16S rRNA gene amplicon Nanopore sequencing and shotgun metagenomic sequencing the taxonomic and functional structure of the microbial community forming a biofilm on the inner wall of a lab-scale transparent anaerobic biodigester illuminated with natural sunlight. The biofilm was composed of microorganisms involved in the four metabolic processes needed for biogas production. The biofilm proved surprisingly rich in Rhodopseudomonas faecalis, a versatile bacterium able to carry out a photoautotroph metabolism when grown under anaerobic conditions. Our results suggest that this bacterium, able to fix carbon dioxide, could be considered for its use in transparent biogas fermenters in order to contribute to the production of optimized biogas with a higher CH4:CO2 ratio than the biogas produced in regular, opaque digesters. To the best of our knowledge, this is the first study supporting illuminated bioreactors as a new bioprocess for the obtention of biogas enriched in methane.

microbiology↗

bioRxiv2018

Microbial communities of the Mediterranean rocky coast: ecology and biotechnological potential

Microbial communities from harsh environments hold great promise as sources of biotechnologically-relevant strains. In the present work, we have deeply characterized the microorganisms from three different rocky locations of the Mediterranean coast, an environment characterised by being subjected to harsh conditions such as high levels of irradiation and large temperature and salinity fluctuations. Through culture-dependent and culture-independent techniques, we have retrieved a complete view of the ecology and functional aspects of these communities and assessed the biotechnological potential of the cultivable microorganisms. A culture-independent approach through high-throughput 16S rRNA amplicon sequencing revealed that all three locations display very similar microbial communities, suggesting that there is a stable community associated to the sampled region, with Stanieria cyanosphaera, Rubrobacter sp. and the families Flammeovirgaceae, Phyllobacteriaceae, Rhodobacteraceae and Trueperaceae being the most abundant taxa. Furthermore, shotgun metagenomic sequencing results were in concordance with the high-thoughput 16S rRNA, and allowed a description of the eukaryotic and archaeal members of the community, which were abundant in Ascomycota and halotolerant archaea, respectively. The culture-dependent approach yielded a collection of 100 isolates (mainly pigmented), out of which 12 displayed high antioxidant activities, as proved with two in vitro (hydrogen peroxide and DPPH) and an in vivo (model organism C. elegans) assays.

microbiology↗

bioRxiv2018

Bioprospecting the solar panel microbiome: high-throughput screening for antioxidant bacteria in a Caenorhabditis elegans model

Microbial communities that are exposed to sunlight typically share a series of adaptations to deal with the radiation they are exposed to, including efficient DNA repair systems, pigment production and protection against oxidative stress, which makes these environments good candidates for the search of novel antioxidant microorganisms. In this research project, we isolated potential antioxidant pigmented bacteria from a dry and highly-irradiated extreme environment: solar panels. High-throughput in vivo assays using Caenorhabiditis elegans as an experimental model demonstrated the high antioxidant and ultraviolet-protection properties of these bacterial isolates that proved to be rich in carotenoids. Our results suggest that solar panels harbor a microbial community that includes strains with potential applications as antioxidants.

microbiology↗

bioRxiv2018

Liquid co-substrates repower sewage microbiomes

A range of parameters are known to shape the methanogenic communities of biogas-producing digesters and to strongly influence the amount of biogas produced. In this work, liquid and solid fractions of grass biomass were used separately for semicontinuous batch methanation using sewage sludge as seed sludge. During 6 months of incubation, the amount of input COD was increased gradually, and the underlying methanogenic microbiome was assessed by means of microscopy-based automated cell counting and full-length 16S rRNA high-throughput sequencing. In this sense, we prove for the first time the suitability of the ONTMinION platform as a monitoring tool for anaerobic digestion systems. According to our results, solid-fed batches were highly unstable at higher COD input concentrations, and kept Methanosaeta spp. typically associated to sewage sludge-as the majoritary methanogenic archaea. In contrast, liquid-fed batches developed a more stable microbiome, proved enriched in Methanosarcina spp, and resulted in higher methanogenic yield. This work demonstrates the high repowering potential of microbiomes from sewage sludge digesters, and highlight the effectiveness of liquefied substrates for increasing biogas productivity in anaerobic digestions.

microbiology↗

bioRxiv2018

Thermoelectric Heat Exchange and Growth Regulation in a Continuous Yeast Culture

We have designed a thermoelectric heat exchanger (TEHE) for microbial fermentations, able to control the temperature of a microbial continuous culture, and produce electric power. The system proved able to stably maintain both the temperature and the optical density of the culture during the exponential, highly productive phase.

microbiology↗

bioRxiv2017

Warm and wet: robust lipase-producing bacteria from the indoor environment

Lipases are key biocatalysts with important biotechnological applications. With the aim of isolating robust lipolytic microbial strains, we have analyzed the bacterial communities inhabiting two domestic extreme environments: a thermophilic sauna and a dishwasher filter. Scanning electron microscopy revealed biofilm-forming and scattered microorganisms in the sauna and dishwasher sample, respectively. A culture-independent approach based on 16S rRNA analysis indicated a high abundance of Proteobacteria in the sauna sample; and, a large amount of Proteobacteria, Firmicutes, Cyanobacteria and Actinobacteria in the dishwasher filter. With a culture-dependent approach, we isolated 48 bacterial strains, screened their lipolytic activities on media with tributyrin as the main carbon source, and finally selected five isolates for further characterization. These strains, all of them identified as members of the genus Bacillus, displayed optimum lipolytic peaks at pH 6.5 and with 1-2% NaCl, and the activity proved very robust at a wide range of pH (up to 11.5) and added NaCl concentrations (up to 4%). The thermal, pH and salt robustness of the selected isolates is a valuable attribute for these strains, which are promising as highly tolerant biodetergents. To our knowledge, this is the first report regarding the isolation from an indoor environment of Bacillus strains with a high potential for industry.

microbiology↗